The present invention relates to a process for the production of a protein concentrate containing immunological factors preferably of lactic origin and to a specific immunoglobulin population. More particularly, this invention relates to active immunoglobulins exhibiting specificity against the microorganism, Helicobacter pylori (formerly referred to in the literature as Campylobacter pylori) and the use of this protein in the management of Helicobacter pylori colonization in the gastrointestinal (GI) tract. More specifically, the present invention relates to the injection of lactating mammals with Helicobacter pylori, subsequent isolation and concentration of antibodies from the colostrum (or milk) produced by the immunized mammals and use of this concentrate, by way of enteral ingestion, in reducing the infectivity of Helicobacter pylori resident in the gastrointestinal tract. This protein (immunoglobulin) concentrate is useful in the treatment of pathological sequela associated with Helicobacter pylori colonization of the GI tract including gastritis and peptic ulcer disease.
In general, the prior art discloses the introduction and use of immunological factors of lactic origin into-dietetic products for newborn babies and infants. The oral ingestion of these dietetic products being intended to enable these immunological factors to be utilized in the development of protection against the consequences of microbial infection within the GI tract.
U.S. Pat. Nos. 3,992,521 and 3,984,539 disclose a process for obtaining an immune product containing antibodies from the serum of a horse or cow and the immunoglobulin product itself. These patents do not suggest nor disclose the specific immunoglobulin of the present invention, nor its method of production and isolation, nor its intended method of use for the treatment of Helicobacter pylori induced gastritis.
U.S. Pat. No. 3,128,230 discloses a method for producing antibodies which consists of injecting a lactating mammal with a mixture of killed microorganisms and isolating the antibodies from serum or milk. This patent does not suggest nor disclose that Helicobacter pylori can induce an antibody response nor the specific method of treatment employed in this invention.
British Patent No: 1,573,995 discloses and claims a process for the production and isolation of immunoglobulins exhibiting specificity against Escherichia coli. This patent does not suggest that microorganisms other than E. coli are useful. In similar fashion, the following references disclose the same type of process: 1) H. Hilpert, et al., Proceedings of the 13th Symposium Swedish Nutritional Foundation; and 2) C. Mietens, et al., European J. Pediatrics, 132, 239-252, (1979).
Ebina and colleagues disclose the immunization of cows with human rotavirus and the isolation of immunoglobulin to the virus from the milk of cows. This immunoglobulin was orally administered to children and was found to reduce the frequency of the outbreak of diarrhea. See Ebina, et al. The Lancet (Oct. 29, 1983), 1029-1030, (1983); and Ebina, et al., Med. Microbioi. Immunol., 174, 177-185, (1985). These references do not suggest nor disclose that immunoglobulins to Helicobacter pylori would be useful in the management Helicobacter pylori induced gastritis.
U.S. Pat. No. 4,051,231 discloses the isolation of immunoglobulins from the milk of vaccinated cows by coagulating the milk, recovering the lactoserum (whey) and selectively precipitating-the immunoglobulins with ammonium sulfate, followed by dialysis against water, filtration and drying. Seroprotection tests demonstrated that the protein concentrates of U.S. Pat. No. 4,051,231 provided local passive immunity in the intestine without resorption and without any significant loss of activity in the digestive tract, thereby providing generalized passive protection against certain enteropathogenic bacteria and/or viruses. This patent does not suggest nor disclose that such antibodies could serve to modify the course of Helicobacter pylori induced gastritis.
Much has been published regarding Helicobacter pylori itself. Helicobacter pylori is approximately 0.85 um in diameter with an average length of 2.9 um. The microorganism has a smooth coat and four to six polar flagella which are sheathed and have bulbous ends. In fresh cultures this organism appears as a slender, curved Gram-negative rod. Helicobacter pylori is readily distinguished from other gastric bacteria and spirochaetes by the absence of axial filaments in its flagella. Furthermore, optimum growth conditions for Helicobacter pylori are unusual and help to set it apart from other enteropathogens. For example, Helicobacter pylori requires a microaerophilic gas environment (i.e. low oxygen content) to sustain growth. Helicobacter pylori appears to tolerate a wide range of local pH conditions and is relatively resistant to acid conditions. It is believed that this resistance is due in part to the organism's outer protein structure which contains urease in large amounts resulting in the cleavage of urea naturally present in gastric fluid and hence, the formation of a buffering ammonia layer immediately around the organism.
Although a number of spiral bacteria inhabit the mouth and lower intestinal tract of all mammals, what distinguishes Helicobacter pylori is the observation that it is localized almost exclusively to the luminal mucosal surface of the stomach and duodenum and generally is found deep within the gastric pits.
It is the combination of the unusual growth requirements and intestinal location which makes eradication and treatment of Helicobacter pylori so difficult. The ideal antimicrobial drug suitable for the successful treatment of Helicobacter pylori associated gastritis should exhibit local activity, be stable at low pH values and should be able to readily penetrate the gastric mucosa. These desirable properties of an antimicrobial are not easily accomplished and thus, satisfactory treatment of Helicobacter pylori with antimicrobials has yet to be accomplished.
The development of an agent which is effective in the management of Helicobacter pylori induced gastritis would fulfill a long felt need. There is an emerging consensus in the field of gastroenterology that Helicobacter pylori is a major contributing-factor in the development of gastritis and septic ulcer disease. Specifically, the following reference is useful in establishing the background of the present invention: Campylobacter pylori, E. A. J. Rauns and G. N. J. Tytgat, Editors, Adis Press Intntl. (1989).
In general, this reference discloses, at pages 138-139, the role of Helicobacter pylori in the development of gastritis and peptic ulcer disease. The key evidence in support of Helicobacter pylori etiology in these conditions is based on the observation, at pages 89-103, that elimination of Helicobacter pylori from the stomach through the use of antibiotics and/or bismuth compounds leads to a remission of the gastric disease.
Presently, the main therapies employed in the treatment of chronic active gastritis and peptic ulcer disease include the histamine H2-receptor antagonists, bismuth compounds, and antibiotics. However, it is generally accepted that all currently used treatment modalitites are clinically inadequate since post-treatment relapse rates remain unacceptably high. In addition, several of these therapies are accompanied by significant side effects. For example, effective antibiotic treatment of Helicobacter pylori infections requires treatment over an extended duration (4-6 weeks) and results in the induction of diarrhea and intestinal discomfort. The bismuth compounds are also known to have a number of significant undesirable side effects.
To date, the preferred treatment has been dominated by the use of H2-antagonists which result in the suppression of acid and pepsin secretion; however, post treatment relapse rates are extremely high. Since symptomatic relief and ulcer healing are the primary aim of treatment, without indefinite maintenance therapy, it is becoming increasingly apparent that a mucosal "protective agent" having antimicrobial activity against Helicobacter pylori, is desirable.
Thus, the medical community has a need for a protective agent which can be readily utilized in pharmaceutical and/or nutritional formulations. The present invention fulfills that need through the discovery that enteral ingestion of immunoglobulins derived from lactating mammals immunized with Helicobacter pylori provides such protection.
The prior art fails to suggest, disclose or contemplate the instant discovery which is, in part, the use of antibodies (immunoglobulin) in the treatment of Helicobacter pylori infection of the gastric mucosa and to the antibodies themselves.
Lactile secretion derived antibodies obtained from cows immunized with Helicobacter pylori will provide numerous advantages over other methods of immunoglobulin production. The advantages include quantity, ease and reproducibility of immunoglobulin isolation, ease of product preparation and significant cost savings as compared to antibody and product preparation based on other isolation methods.
One aspect of the present invention relates to a method for producing a milk based product having high immunological specific activity against Helicobacter pylori.
Another further aspect of this invention is the specific immunoglobulin itself which is produced according to the disclosed method.
A further aspect of the present invention relates to a method for treating mammals in order to produce milk having immunological components which provide protection against Helicobacter pylori to subjects imbibing same.
A further aspect of this invention is the use of these specific antibodies (immunoglobulins) in the treatment of Helicobacter pylori induced gastritis.